To identify whether microtubules are involved in the maintenance of vacuolar morphology, we treated Saccharomyces cerevisiae with nocodazole and methyl benzimidazole-2-yl-carbamate, drugs which inhibit the polymerization of microtubules. Treated cells arrest with a single large bud in the G2/prophase portion of the cell cycle. Labeling the vacuole with either quinacrine or FITC-dextran revealed vacuole fragmentation that was not found in untreated cells or in cells arrested in G2 by unrelated means. A drug-resistant mutant in beta tubulin does not show vacuolar fragmentation when treated with drug. We propose that microtubules are involved in the regulation of vacuole morphology.